Temperature Control System for Biochemical Reactions in Microchip-Based Devices

IntroductionIn recent years,the development of miniaturizedsystems for chemical and biochemical reactions hasrapidly progressed[1] .Most of the microdevices forconducting these reactions are made of silicon orglass using conventional microfabrication methodsused in the microelectronic industry or bymodifying the processing technologies for microelectromechanical systems ( MEMS) . Severalresearch groups have developed polymerase chainreactions ( PCR) chips with various features toperform sim…     

PCR产物计算公式的推导与修正

从PCR放大产物的形成和组成特点入手，推导出基因组特异DNA序列扩增倍数的理论计算公式，即Yn=An（1＋X）^n，其中Y表示放大后的DNA片断拷贝数，x表示各反应周期的平均放大效率，n代表循环次数，An为反应周期系数。     

人rabl3基因克隆和表达

以人的胚胎ｃＤＮＡ为模板，用ＰＣＲ方法筛选获得ｒａｂ１３基因，并克隆到真核表达载体ｐｃＤＮＡ３和原核表达载体ｐＧＥＸ、ｐＥＴ－１５ｂ和ｐＭＡＬ－ｃ２中，把ｒａｂ１３基因转染入牛肾上腺毛细血管内皮细胞（ＢＣＥ细胞）中，免疫荧光染色显示Ｒａｂ１３定位于ＢＣＥ细胞胞质内膜性细胞器上，在为ｒａｂ１３基因对膜通道调节功能的进一步研究打下了基础。     

实时定量PCR鉴定转基因作物纯合体

以水稻内标准基因磷脂酶D基因(phospholipase D,PLD)和转基因水稻TT51-1特异性旁侧序列为检测靶标,对单株转基因水稻的种子播种后长出的单株进行了荧光定量PCR,以其内标准基因和旁侧序列Ct值的差值判断了植株的纯合体,并用标准曲线计算了转基因含量,证实了此法的可靠性,说明此法用于鉴定植株的纯合体简便准确.     

脉胞菌hH3v基因的初步研究及原核表达载体构建

根据植物表达载体pET52（b）多克隆位点和脉胞菌着丝粒结构相关蛋白CenH3的编码基因hH3v序列，设计引物，通过PCR扩增将基因两端加上KpnⅠ和SacⅠ酶切位点，经双酶切后将hH3v插入表达载体pET52（b）中，利用热激法转化大肠杆菌Top10感受态细胞，在选择培养基上挑取单菌落培养，经PCR检测，表明目的基N原核表达载体构建成功，为着丝粒结构进一步的研究提供了基础．     

Enhanced activity of yqhD oxidoreductase in synthesis of 1,3-propanediol by error-prone PCR

yqhD oxidoreductase was determined to be an NADP-dependent dehydrogenase, and was more active toward 3-HPA when compared to 1,3-propanediol oxidoreductase. To further improve enzyme activity towards 3-hydroxypropionaldehyde （3-HPA）, error-prone PCR was implemented to mutant yqhD gene. Two mutants, D99QN147H and Q202A with increased catalytic and affinity efficiency, were obtained after one round of error-prone polymerase chain reaction. And the catalytic efficiency of the mutant D99QN147H was up to 4-fold greater than the wild enzyme （0.0375 min^-1 mM^-1 vs. 0.0078 min^-1 mM^-1）. The recombined strain containing pET28yqhD D99QN147H yielded 28 g L^-1 of 1,3-propanediol in the fed-batch LB cultures （1 L volume） with an initial 3-HPA concentration of 40 g L^-1, which was higher than the 21 and 17 g L^-1 of 1,3-propanediol from the mutant Q202A and the wild-type, respectively. Except for propionaldehyde, the optimal mutant D99QN147H also exhibited higher activity on a range of substituted aldehydes than the wildtype.     

原位多聚酶链式反应（In Situ PCR）技术的应用和发展

原位ＰＣＲ是一项新的分析子技术，它结合了杂交技术和ＰＣＲ技术的优点，具有高度的敏感性，特异性，并能进行精确的定位，在研究工作和临床诊断上有广阔的应用前景。     

几种动物RAPD指纹图谱反应条件的分析

用随机排列碱基顺序的寡核苷酸引物，对蚕、鱼、蛇、家猪等动物的基因组ＤＮＡ扩增，获得了这些动物的随机扩增多态ＤＮＡ指纹图谱，对影响扩增结果的几个因素进行了分析，对动物随机扩增多态ＤＮＡ反应的适宜条件进行了讨论。     

用加端PCR技术改造h－IGF－1cDNA

用ＤＮＡ合成仪合成了分别带有ＰｓｔＩ位点和ＳａｌＩ位点及终止密码子的２个用于扩增ｈＩＧＦ－１ｃＤＮＡ的ＰＣＲ引物．利用合成的引物，７００ｂｐ长的ｈＩＧＦ－１ｃＤＮＡ模板和Ｔａｑ聚合酶进行ＰＣＲ扩增．扩增产物经电泳鉴定后克隆进Ｍ１３ｍｐ１８载体，进行核苷酸序列分析．结果显示：ＰＣＲ产物含已发表的ｈＩＧＦ－１成熟蛋白的编码序列和５＇端的ＰＳｔＩ位点及３＇端的ＳａｉＩ位点及终止密码ＴＡＧ．用加端ＰＣＲ技术成功地扩增和改造了ｈＩＧＦ－１的编码序列．     

Studies inin situ PCR detected by the BrdU antibody technique

Using the BrdU antibody technique followed by an immuno-chemical staining (BAT), the amplification of DNA fragments specific to human Y chromosome on cell specimen slides was efficiently detected. Whether direct BrdU incorporation into PCR products orin situ hybridization with PCR products on slides, the amplified target DNA fragments of specimen were visualized by BAT under the microscope. The availability of BAT and differences in the sensitivity and efficiency between BAT and dig-11-dUTP labeling in cellin situ PCR were discussed. Zhang Xiyuan: born in Oct. 1935, Professor, Current research interest in Cellular and Molecular Biology Supported by the National Natural Science Foundation of China